1996 - v goffin - antidandruffshampoosandthestratumcorneum[retrieved-2014!09!20]
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Journal of Dermatolog ical Treatment (1996) 7. 21 5 - 2 18
@ 1996 Journal
of
Dermatological Treatment.Al l rights resewed 0954 6634
Antidandruff shampoos and the stratum corneum
V Goffin, C Piirard-Franchimont G E Piirard
Belgian SSTC Research Center
5596,
Department
of
Dermatopathology. Uiriversiry
of
Liige. Liige. Belgium
The clinical efficacy of antidandruff shampoos containing
antifungal agents is correlated with their anti-Mdusseziu
activity. Other ingredients
of
the shampoos also affect the
efficacy and tolerability of the treatments. This study was
conducted to quantify the effect of eight proprietary
antidandruff shampoos on the human stratum corneum.
Corneosurfametry and squamometry were used to ra te the
interaction between the shampoos and the stratum
corneurn. Significant differences in mildness were dis-
closed between some test shampoos and SLES-7E0
selected as a mild reference surfactant. The activity
against corneocyte clumping also showed significant
differences among the test products. I t is concluded t hat
the formulation of shampoos for dandruff should not be
overlooked.
( J
Dermatol Treat 1 996) 7: 21
5 -
2 18)
Keywords: Antidandruff sham poos- Stratum corneum
-
Corneosurfametry- Squamometry
Rrceivrd 13th November 1995; accepted 31sr Jul
19%
Materials and methods
Test products
Eight proprietary antidandruff shampoos were used.
They contained one of the following active agents: eco-
nazole nitrate 1
YO,
Pevaryl, Cilag, Berchem, Belgium),
ketoconazole (2%, Nizoral, Janssen Pharmaceutica,
Beerse, Belgium), piroctone olamine (0.5%, Dercos,
Laboratoire D'Anglas, Paris France;
1
Yo
Anatel, La-
boratoire Roche Posay, La Roche Posay, France; 1%
Cystelle, Laboratoire Bailleul, Paris, France), selenium
sulphide (2.5%, Selsun, Abbott, Ottignies, Belgium) and
zinc pyrithione 1
YO,
ead and Shoulders, Procter and
Gamble, Strombeek Bever, Belgium; 1YO, inkan, Le-
derle Cyanamid. Louvain-la-Neuve, Belgium). Sodium
laureth sulfate EO 7: 1 (SLES-7EO) as a 2% aqueous
solution served as the reference product in the squamo-
metric and corneosurfametric tests because it exhibits
only mild ir ri tancy in V ~ V O . ' ~ . ' ~
Introduction
Modem shampoos designed for treating dandruff and
seborrhoeic dermatitis were developed following the
demon stration that the active ingredients are antifungal
agents which inhibit the growth of Malassezia ovalis
(formerly known as Pityrosportrni ov ale) . -3The ultimate
goal of these shampoo s is to remove scales and to alter
the corneocyte binding and multiplication of this re-
sident lipophilic yeast. Removing the scales is important
both for the immediate cosmetic effect and for blocking
the cyclic process during which the yeast accumulates,
releases proinflammatory mediators and activates an
immune r e~ po ns e, ~ hich in turn disturbs the pro-
liferation and maturation of the epidermis. The resulting
thick and porous horny layer is the niche in which the
yeast continues to flourish. The effect of shampoos on
intercorneocyte cohesion is largely unknown.
Several investigational methods have been developed
to rate desquamation and the potency of squamolytic
agents6-' Among them, squamometric and corneo-
surfametric bioassays give insight into both the cohe-
siveness and structural changes occurring in
co rne~cy tes . ' .~hese methods are particularly suited for
evaluating the interaction of surfactants with the hum an
stratum corneum.I0-l3
The aim of this study was to rate the effects of anti-
dandruff shampoos on the
stratum corneum. Non-
invasive squamometric and corneosurfametric tests were
conducted to evaluate the effects of eight proprietary
shampoos on the stratum corneum.
Correspondence: Prof G Pierird, DepJrrment of Dermatopathology.
CHU Sari Ti lman, 8-4000, iege, Belgium, Tel. 32-41-662408; Fax.
+ 32-41 -662976
Squamometric in vivo bioassay
A grou p of 12 volunteers, aged 23-47 years, entered a
double-blind within-subject comparison after signing an
informed consent. The iterative short-term patch pro-
tocol consisted of occlusive patches applied three times
for 30 min
at
2-h intervals to the volar forearms.'.' Each
time, a fixed amount (0.15 ml) of water, or 2% solutions
of the shampoos or SLES-7E0 was deposited into
19-mm Hill Top chambers. Two hours after the final
application of the test products, clear adhesive-coated
discs (D-Squame, Cuderm Corporation, Dallas, TX)
were placed under controlled pressure
(1
10 g/cmz) for
5 s
onto each test site. After careful removal, they were
stained for 1 min with
a
solution of basic fuchsin-
toluidine blue (PMS, polychrome multiple stain, De-
lasco, Council Bluff, Iowa). The discs were evaluated
colorimetrically as previously described for the squa-
mometric method. - Da ta are expressed as colour
differentials, AE*ab, between the material collected
from each test site and from the water-treated site. This
variable is referred to as the differential squamometry
index (D-SQMI). In addition, the same samples were
examined under the microscope for rating both the in-
tensity of the corneocyte PMS staining and the pattern
of intercorneocyte cohesion (Table I, Figure 1).
Corneosurfametric
ex
vivo bioassay
Th e 12 volunteers, who part icipated in the in vivo
squamometry study also participated in this corneo-
surfametric bioassay study. The method entailed col-
lection of cyanoacrylate skin surface strippings (CSSS)
from healthy individuals and a 2-h contact time
with
a
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216 V
Go ffn et al
Table
I
Interval scales for microscopic grading o f D-Squame
adhesive-coated discs
Score Preilorninunt pat tern
Staining intensity
0 No
staining
1
Discrete between corneocyta
2
Spotted
on
corneocytes
3
4
Strong
in
most corneocytes
0
1
2
3
4
Sinnleltorn corneocvtes
Strong in single and/or clustered corneocyta
intercorneocyte loosening
Large and uniform sheet of corneocytes
Large clusters. few single corneocyta
Small clusters, some single corneocytes
Disrupted clusters, many single corneocytes
2% solution of the test
product^.'^-^'
After staining the
samples with PMS for 3 min, rinsing and air drying,
reflectance colorimetry (Chroma Meter, Minolta, Osa-
ka, Jap an) was used to derive values of L* nd Chroma
C .
The colour differential, AE*ab, was calculated
be
tween each sample and a reference CSSS sprayed with
water only. This value
is
referred to as the comeo-
surfametry index (CSMI) which increases with the irri-
tation potential o f the skin-cleansing produ ct being
Statistical analysis
The distribution of each numerical variable CSMI, -
SQMI) was characterized by the mean (M), SD and
coefficient of variatio n
V=
o2 SD/M). Differences be
tween the test and reference products were tested with
the paired Student's t-test. The Wilcoxon statistical
processing of ordinal scales signed ranks test was used
for the microscopic grading of the D-Squames. Re-
gression analysis was applied to determine the re-
lationship between variables. The linear, logarithmic,
exponential and power models were tested with calcu-
lation of the coefficient of correlation. All results were
considered to be significant at the 5 critical level
P< .05).
Results
The results of the squamometry and the comeosurfa-
metry are presented in Table 11.
A
significant linear
a
b
Figure
1
Microscopic aspect of squamometry samples after PMS
staining
see
Table
I).
(a) Example
of
score
1
in both staining
intensity
and
intercorneocyte
loosening.
b) Example of
score
3
in
staining intensity and score I in intercorneocyte loosening. (c)
Example of scon 3 in both staining intensity and intercorneocyte
loosening.
Table Corneosurfam etry and squamom etry. Colorimetric evaluation of
the
corneosurfametry index CSMI) nd of the differential
squamometry index (D-SQMI). Microscopic evaluation
of
the corneocyte dye binding and of
the
pattern
of
intercorneocyte cohesion
on D-
Squames
Brand name Antifungal agent CSMI
Wab)
D-SQMI A F a b ) D-ve binding Corneocyte loosening
Pevaryl Econazole nitrate
1
32.8f .5'
15.3 k5 .3 2.1 f0 .6
Nizoral
Ketoconazole 2
25.5f 6.3
12.6f4.7. 1.8f 0.6 1.7f0.5'
Dercos Piroctone olamine 0.5 28.4k7.7
14.9k8.5 1.7f0.9 1.6f0.3'
Anatel Piroctone olamine
1YO 24.6k8.3
9.2f
5.9
1.3f0.8 0.5f0 .3 .
Cystelle Piroctone olamine
1
37.8
f 4.2'
16.8f8.4 2.4f
.5.
0 .9 f 0 .8
Selsun Selenium sulphide
2.5
48.3f15.8'
l 7 . 9 k
8.3
2.6f0 .5* 1 .8k1.2
Head and Shoulders Zinc pyrithione
1%
27.7f9 .9 13.6f5 .4
1.7
f 0 . 7 1 . 1 f l . 3
Zinkan
Zinc pyrithione
1
24.5f .2
17. 45 7.1 2.0k0.9 1.3k1.5
Reference product SLES-7EO
23.1k6.8 16.3f .4 1.8f0.3 1.3f0 .4
Shampoo Corneosurfametry Squamome t r y
0 .9 f 0 .6
*Significantly different
(p
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An t i d a n d r u f f Fharnpoos
and
Ihe
ctraturn
corneum
217
correlation
( r = 0 . 5 4 . ~ 0 . 0 5 )
as found between C SM I
and D-SQMI values of all the shampoos combined. The
interindividual coefficients of variation were lower for
CSMI than for D-SQMI, ranging from
25
to
38%
and
from 3 5 to 65%, respectively. CSMI was linearly cor-
related
r =
0.78.
Pc0.01)
ith the rating of dye binding,
but was unrelated
( r = O . 2 3 ,
NS) to the rating of cor-
neocyte loosening. A power relationship ( r
= 0.62,
P < .05) was found between D-SQMI and the intensity
of corneocyte loosening.
Some shampoos showed significant differences from
the reference produc t
SLES-7E0
(Table 11). Th e mildest
shampoos showing prominent anticlumping activity
(Anatel, Dercos, Nizoral) were identified by b oth low D-
SQMI values and high corneocyte loosening in the
squamometric test. Two shampoos (Cystelle, Selsun)
were significantly more irritan t than
SLES-7E0
because
they showed increased CSMI values and increased dye
binding. The other shampoos (Head and Shoulders,
Pevaryl, Zinkan) formed an intermediate category
of
products which globally resembled the reference
sur-
factant SLES-7E0.
Discussion
The pivotal role of M . ovalis in the development of
dandruff is generally acknowledged. It has been shown
that the activity of antidandruff shampoos of the anti-
fungal class is primarily linked to their activity against
Mafusse~ia.'
n the global clinical evaluation, the other
ingredients of sham poos m ay interfere with the efficacy
an d tolerability of the treatment. One aspect is related to
the ability to remove a nd disrupt clumps of corneocytes
without inducing irritation of the scalp. Surfactants
present in shampoos are interfacially active substances
which interact with the stratum corneum. The mechan-
ism(s) whereby surfactants elicit undesirable effects is
not fully understood.16 Many techniques for the esti-
mation of the potential irr itancy of surfactants have
been described over the past
30
years. Traditional eva-
luations usually entail assessing the degree of erythem a,
dryness and scaling. -19 M os t of the methods categorize
products into a few broad categories. However, they
lack the sensitivity to distinguish products within a
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